What are your recommendations for the best way to dilute and store my TaqMan® probe?

Product FAQ

Answer

The probe should be diluted in TE buffer [10 mM Tris-HCl, pH 8.0 (at 25°C), 1 mM EDTA]. We recommend storing your probes, both stocks and working solutions, at -20°C in aliquots. This is to cut down on the number of times the probe is subject to freeze-thaws. For detailed information on diluting primers and probes, please refer to the tutorial entitled "Reconstituting and Diluting Primers and TaqMan® Probes". You can find it on our website by entering this title in the main search field.

Answer Id: E1129

Was this answer helpful?

Yes
No
12a80cb6ae33839ee36b79168b7b308a_FAQ

When using the TaqMan® Gene Signature Plates, should I mix the reactions after adding the master mix and template to the wells containing lyophilized primers and probe?

Product FAQ

Answer

No, but it is recommended that the plate be centrifuged to draw down the liquid to the bottom of the wells.

Answer Id: E1370

Was this answer helpful?

Yes
No
c6fa0858af353629b94e322efb5117f3_FAQ

What are the key features of a TaqMan® MGB probe?

Product FAQ

Answer

TaqMan® MGB probes have a nonfluorescent quencher at the 3´ end and a minor groove binder at the 3´ end that increases the melting temperature (Tm) of probe, allowing the use of shorter probes.  For more information on MGB probes, please consult the following manual: "Primer Express v1.5 and TaqMan® MGB Probes for Allelic Discrimination: All PCR Instruments: User Bulletin." You can find a copy on our website by entering this title in the main search box.

Answer Id: E1371

Was this answer helpful?

Yes
No
bd9ac49d36e1a373a4d3d94200ceafd7_FAQ

Can TaqMan® Gene Expression Assays be used in a multiplex reaction?

Product FAQ

Answer

These assays have not been tested to run in a multiplex environment. In addition, each assay is pre-formulated with a 6-FAM™ reporter dye, so it is not possible to incorporate two different TaqMan® Gene Expression Assays into a multiplex reaction. If you would like to multiplex TaqMan® Gene Expression Assays with an endogenous control gene, our TaqMan® Endogenous Control product line offers some commonly used housekeeping genes for human, mouse, rat, and other species, and one assay that will detect 18S rRNA from all eukaryotic organisms. These assays are available with a VIC® reporter dye and in primer-limited formulations suitable for multiplexing. To see the list of these assays, please type "TaqMan Endogenous Control" in the web search form to search and it will lead you to the product page. For information about optimizing a multiplex gene expression experiment, please see "User Bulletin #5: Multiplex PCR with TaqMan® VIC® Probes (P/N 4306236B)". You can get the document by using "User Bulletin #5' as a keyword to search our web site. This User Bulletin was written for use on the ABI PRISM® 7700, but the concepts are valid for all of our real-time PCR instruments.

Answer Id: E1836

Was this answer helpful?

Yes
No
a6584b5919f1dbdd489a05ef029a3b49_FAQ

Do you offer a human beta actin probe with a reporter dye other than FAM™ dye?

Product FAQ

Answer

We do offer two VIC® labeled beta actin probes as part of TaqMan® Endogenous Controls:

1) Human beta actin (VIC®/ TAMRA™ Probe, Primer Limited) Cat. No. 4310881E
2) Human beta actin (VIC®/ MGB Probe, Primer Limited) Cat. No. 4326315E

Please consult our Real-Time PCR Applications page for more information.

Answer Id: E1571

Was this answer helpful?

Yes
No
872d709ac98afaae307c25497785d7bc_FAQ

Can I use the TaqMan® β-actin Detection Reagents (Cat. No. 401846) as an endogenous control for my gene expression studies?

Product FAQ

Answer

The TaqMan® β-actin Detection Reagents were designed to amplify genomic DNA, and thus are not recommended for use as an endogenous control for gene expression studies. However, we do offer other TaqMan® assay reagents that were specifically designed for that application. Two examples are: Human ACTB (Beta Actin) Endogenous Control, Cat. No. 4310881E (TAMRA™ probe) or Cat. No. 4326315E (MGB probe).

There are many factors to consider when validating a specific endogenous control. For additional information on designing gene expression assays and controls, please refer to the following documents: "Protocol: TaqMan® Human Endogenous Control Plate", and User Bulletin #2: "Relative Quantitation of Gene Expression". You can find copies of these documents on our website by entering the titles in the main Search field.

Answer Id: E1780

Was this answer helpful?

Yes
No
bbe98bb766aa4da1b71b4506c3f08552_FAQ